Natural, anti-bacterial, anti-virus, anti-herpes cream

ABSTRACT

Therapeutic compositions made from the herb  Tribulus terrestris  and methods of making and using the same are provided. The therapeutic compositions include an enriched extract having an increased spirostanol saponin content that is prepared from the harvested  Tribulus terrestris  L. The enriched extract is prepared using discrete hydrolysis, separation and enrichment steps. The resulting therapeutic may be combined with a cream base and is useful for treating bacterial, fungal, and viral infections, particularly gynecologic infections. This product was also found to be very successful in suppository form for the treatment of vulvo-vaginal, vulvo-hemorrhoidal and colonic conditions.

CROSS-REFERENCES TO RELATED APPLICATIONS

[0001] This patent application claims priority from U.S. Pat. No.6,343,258 B1 filed Aug. 13, 1999 for METHOD FOR TESTING FOR READINESS OFHARVESTING OF TRIBULUS TERRESTRIS L. HAVING HIGH STERIODAL SAPONINCONTENT and is a continuation-in-part of U.S. patent application Ser.No. 10/016,085 filed Dec. 12, 2001 for NATURAL, ANTI-BACTERIAL,ANTI-VIRUS, ANTI-HERPES CREAM, which are incorporated herein by thisreference thereto.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] This invention relates to the field of therapeutic compounds forthe treatment of diseases of the skin and mucosal tissue. In particularthe present invention relates to creams, ointments, eye and ear dropsmade from the herb Tribulus Terrestris that have a high content ofspirostanol-type steroidal saponins and are useful in the treatment ofconditions related to viral, fungal, yeast, parasitic and bacterialinfections, particularly genital, skin (including acne) and eye and earinfections.

[0004] 2. Description of the Related Art

[0005]Tribulus Terrestris, commonly known as “Puncture Vine” or Caltropfruit, is an herb that has been used for centuries in Europe for hormoneinsufficiency in men and women. It has also been used in the treatmentof liver, kidney and urinary tract disease. In recent years TribulusTerrestris has been touted as a dietary supplement for improvingathletic performance. It has been discovered that ingestion of Tribulusterrestris significantly elevates the level of several hormones:Testosterone; Luteinizing Hormone; Follicle Stimulating Hormone; andEstradiol. Clinical studies on Tribulus, conducted at the ChemicalPharmaceutical Institute in Sofia, Bulgaria, showed improvedreproductive functions, including increased sperm production andTestosterone levels in men. Among women, Tribulus terrestris L.increased the concentration of hormones including Estradiol, withTestosterone being very slightly influenced, thereby improvingreproductive function, libido and ovulation.

[0006] The active components of the Tribulus terrestris L. plant alsohave a stimulating effect on the immune, sexual and reproductivesystems, leading to improved muscle building, stamina and endurance.Other positive changes observed in a number of cases were a reduction incholesterol, enhanced mood and well-being. No adverse effects to thecentral nervous or cardiovascular systems were noted in any of theclinical studies.

[0007] The Tribulus plant is also of interest due to the increasingclinical importance of emerging pathogens as well as drug-resistantmicroorganisms, which lend additional urgency to identify novel, activecompounds. Currently available drugs for the treatment of fungalinfections include amphotericin B (a macrolide polyene), which interactswith fungal membrane sterols, flucytosine (a fluoropyrimidine), whichinterferes with fungal protein and DNA biosynthesis, and a variety ofazoles (e.g. ketoconazole, itraconazole, and fluconazole) that inhibitfungal membrane-sterol biosynthesis (see e.g. Alexander, B. D. et al.,Drugs, 1997, 54:657-678). Although amphotericin B has a broad range ofactivity and is viewed as the “gold standard” of antifungal therapy, itsuse is limited due to infusion-related reactions and nephrotoxicity (seeAlexander, B. D. et al., Drugs, 1997, 54:657-678). Flucytosine usage isalso limited due to the development of resistance and its narrowspectrum of activity. Further, the widespread use of azoles is causingthe emergence of clinically-resistant strains of Candida spp. (seeAlexander, B. D. et al., Drugs, 1997, 54:657-678). This problem isparticularly important in treating vaginal infections due to bacteria,parasitis and fungi.

[0008] Other commonly used agents in treating vaginal infections includedimethylalkybenzalkonium choloride, nonylphenololyoxyethylene ornonoxynol-9. The use of these bacteriocidal agents in vaginally insertedsuppositories, creams, foams or the like, however, is also not withoutproblems inasmuch as these agents tend to diminish or destroy thehealthy bacterial flora of the vagina, and cause a tendency to developyeast infections (candidasis). Further, many of the prior artcompositions have a narrow spectrum of action in that they cover onlysome of the microorganisms responsible for a particular condition.

[0009] In this respect, the Tribulus terrestris L. plant has been apromising alternative to existing “chemical” anti-microbials. Tribulusterrestris L. contains saponins, which in plants, are formed from asapogenin core linked with a sugar. The sapogenin, or glycoside, may bea steroid or a triterpene and the sugar moiety may be glucose,galactose, a pentose or a methyl pentose. Samples of Tribulus terrestrisL. extract prepared according to the invention disclosed herein havebeen found to include tigogenine, diosgenin (a steroid sapogenin),desoxydiosgenin astragalin, kampferol (a flavanoid), dioscin (afurostanol that converts to diosgenin and glucose under hydrolysis),protodioscin (a furostanol), harmine (a bioflavanoide), linoleic acid,galactoolysaccharide, protogracillin and kampferol biofavanoidecompounds. As with many secondary plant compounds, the concentration ofsaponins within a plant will vary depending upon the identity of theplant, the source of saponin within the plant (e.g. seed, fruit, leaves)and the conditions under which the plant is grown. The method ofextraction will also affect the concentration of saponins. Generally,however, saponins are present in conventional extracts at about 15 to 25percent by weight and average at about 20% by weight.

[0010] Saponins are also found in soybeans, alfalfa, and ginseng havebeen studied extensively for their effect of lowering cholesterol. Forexample, U.S. Pat. No. 4,242,502 issued to Malinow et al (1980) relatesto the use of saponins to inhibit cholesterol absorption. According tothis reference, modification of the oligosaccharide portion of thesaponin by hydrolysis under mild acid conditions affects the saponins'ability to affect cholesterol absorption. More particularly, when thesugar moiety of the saponin molecule is removed, i.e., the glycosidiclinkage is cleaved, it was found that the cholesterol is no longerremoved. Therefore, hyrolysis of saponin in the stomach or intestine isan important factor in considering the efficacy of saponin-containingtherapeutic compound, for example, in the treatment ofhypercholesterolemia. To this effect, U.S. Pat. No. 5,760,009 (Allen etal., 1998) discloses a crystalline monohydrate of a spirostanolglycoside that is not degraded in the digestive system and is useful forthe control of hypercholesterolemia or atheroscerosis. Topicalapplication of saponins also presents special consideration fortherapeutic efficacy since not all saponins are topically bio-active.

[0011] The steriodal saponins found in the herb Tribulus terrestris aretwo types; furostanols and spirostanols. Furostanol steroidal saponinsfrom Tribulus terrestris L. are not topically bio-active and must befirst converted to spirostanols to become bioactive, for example, viaenzyme hydrolysis. The instant invention discloses the use of Tribulusterresteris L. compositions having a high spirostanol saponin contentuseful in treating fungal, bacterial viral and parasitic infections andmethods for making the same.

SUMMARY OF THE INVENTION

[0012] The present invention includes pharmacologic compositions derivedfrom the herb Tribulus Terrestris L. that have a high content ofspirostanol-type steroidal saponins and/or balance the ratio offurostanol to spirostanol-type saponins in the resulting composition.The compositions may be used topically, orally and on mucosal tissuesand exhibit a strong anti-bacterial, anti-inflammation, anti-viral, andanti-herpes and hormone balancing effect. They are highly useful intreating vaginal infections, infertility, symptoms of menopause, skindisorders, bacterial infections and viral infections such as herpessimplex I and herpes simplex II, and in some cases may block cancercells from growing. Treatment four times per day with tablets comprisingabout 250 mg of Tribulus terrestris extract was found to significantlyalleviate symptoms associated with the presence of ovarian cysts andbreast cysts. Tribulus terrestris L. compositions, as described herein,may be taken in tablet form to alleviate hormonal irregularitiesassociated with conditions such as female infertility and theundesirable symptoms of menopause (e.g. “hot flashes”).

[0013] The compositions disclosed herein, may be combined withingredients known in the art to provide creams, ointments, oral tabletsand eye and eardrops. For example, the present invention discloses acream made from Tribulus Terrestris L. that has a very stronganti-bacterial, anti-inflammation, anti-virus, anti-herpes effect,represents a great improvement in the fields of dermatology andgynecology and satisfies a long felt need of dermatologists andgynecologists. Additionally, the compounds and methods disclosed hereinare useful in treating cervical wounds, cervical erosion, diabeticwounds, Burger disease, phlebitis, rashes, skin burning, varicose veinsand broken split skin.

[0014] The compositions disclosed herein, include preparations usingmethods comprising isolation, enzyme hydrolysis, and enrichment steps tooptimize the spirostanol content of the resulting therapeuticcomposition. To prepare a preferred composition having optimalspirostanol content, a first extract is prepared by first carrying out,preferably, a low temperature water/alcohol extraction of the TribulusTerrestris L. plant material. A number of factors are important inpreparation of the Tribulus Terrestris L raw material and the extractionprocedure. These include: time of harvesting the Tribulus terrestris L.,part of the herb used, specific geographic area where the herb isgathered, the method of harvesting, and low temperature drying.Adherence to these factors guarantees high steroidal saponin, sapogeninand sterol content of the raw material used for making the extracts andresulting pharmacologic compositions.

[0015] For example, harvesting Tribulus terrestris L. from southernBulgaria between approximately July first and August fifteenth ensuresthat the harvested Tribulus terrestris L. has a high saponin content. Asample of the crop to be harvested can be tested prior to harvestingaccording to the method disclosed in U.S. patent application Ser. No.09/373,812 to further ensure that the Tribulus terrestris L. has optimalsaponin content. Alternately, the crop to be harvested can be testedprior to harvesting according to the method disclosed in U.S. patentapplication Ser. No. 09/373,812, but using a general ratio of one partwater to two and one half parts of dried raw plant material.

[0016] A water/alcohol extraction of the herb Tribulus Terrestris L maythen be performed and the resulting extract concentrated with a vacuumevaporator. To prepare a highly bioactive extract of Tribulus terrestrisL., spirostanol steroidal saponin, sterol and bio-flavanoides, arepreferably isolated from a first Tribulus terrestris L. extract and areblended with a second Tribulus terrestris L. extract. More particularly,in a first step, the first extract is treated to hydrolyze furostanolscontained in the extract to convert them into spirostanols. In a secondstep, the spirostanols in the first extract are isolated and added to asecond prepared Tribulus terrestris L. extract. The resultingcomposition may be mixed with ingredients known in the art to form acream, liquid, tablet or ointment and has been found to be verysuccessful in suppository form for the treatment of bacterial, fungal,yeast, parasitic and bacterial infections.

BRIEF DESCRIPTION OF THE DRAWINGS

[0017]FIG. 1 is a graph showing the importance of vulvo-vaginitissymptoms.

[0018]FIG. 2 is a graph showing how various symptoms improve withtreatment.

[0019]FIG. 3 is a chart of the data used in preparation of FIG. 2.

[0020]FIG. 4 is a graph showing improvement in various symptoms.

[0021]FIG. 5 is a bar chart showing physician assessment of results oftreatment.

[0022] FIGS. 6-1 and 6-2 represent High Pressure Liquid Chromatographydata from a Tribulus Terrestris L. sample as prepared by the methodsdisclosed herein showing the protodioscine peak.

[0023]FIG. 7 illustrates the general formula for a spirostanol.

[0024]FIG. 8 illustrates enzyme hydrolysis of furostanol saponins byB-Glucosidase or acid hydrolysis to from spirostanol saponins.

[0025]FIG. 9 illustrates hydrolysis of Dioscin to Diosgenin.

[0026]FIG. 10 illustrates hydrolysis of furostanols to spirostanols.

[0027]FIG. 11 illustrates changes in the Kupperman index between month 0and month 6 after treatment with Tribulus terrestris L. compositions.

DESCRIPTION OF THE PREFERRED EMBODIMENT(S)

[0028] The detailed description set forth below in connection with theappended drawings is intended as a description of presently preferredembodiments of the invention and is not intended to represent the onlyforms in which the present invention may be constructed and/or utilized.The description sets forth the functions and the sequence of steps forconstructing and operating the invention in connection with theillustrated embodiments. However, it is to be understood that the sameor equivalent functions and sequences may be accomplished by differentembodiments that are also intended to be encompassed within the spiritand scope of the invention.

[0029] The herb Tribulus Terrestris L. is preferably harvested from thesouthern part of Bulgaria where high concentration of steroidalsaponins, sapogenins and sterols in the herb has been confirmed by yearsof analysis. Preferably, the leaves, fruits and stems of the plant areused, without addition of the underground portions of the plant. Priorto harvesting, a sample of the crop is preferably first tested forreadiness of harvesting by a quick qualitative analysis of the steroidalsaponin, sapogenin and sterol content of the plant. For testing, anapproximately 500 mg of harvested and dried (at a temperature of 45 C.)plant material (leaves, fruits and stems) of Tribulus Terrestris L ispreferably used. The dried sample is preferably blended forapproximately 15 minutes in a container with approximately 200 ml water.Upon blending, if the white foam level of the saponins is about 1.5higher than the water level, the plant is ready for harvesting. Once,activity of steroidal saponins is confirmed, the harvesting process ispreferably completed within one week to ensure optimal levels ofsteroidal saponins in the harvested plants.

[0030] Alternately, testing for optimal harvesting time of a sample ofTribulus terrestris L. may be carried out by performing the testingprocedure above, but using dried Tribulus terrestris L. and water at aratio of 2.5 parts dried plant material to 1.0 part water and blendingfor approximately fifteen (15) minutes. The blending time may beincreased as the sample of plant material becomes greater than 500 mg.As in the first method, optimal levels of steroidal saponins in thesample can be confirmed by ascertaining whether the white foam level ofthe saponins is about 1.5 higher than the water level, in which case theplant is ready for harvesting. The optimal time of harvesting Tribulusterrestris L. in Bulgaria is preferably between July 1 and July 30 or,depending on the weather conditions, to as late as August 15.

[0031] Once the plant has been harvested it is preferably stored in adry place or dried immediately. If the herb is dried, it is preferablydone at a low temperature (40° C.) in an oven or at room temperature ina dry, very well ventilated facility. In order to keep the activity ofthe steroidal saponins in the raw material high, the moisture content ispreferably less than 10%. Once harvested, a high speed shredder andmixer may be used to pulverize the plant material to release steroidalsaponins and sapogenins from the plant tissue. Preferably, a mixture ofleaves, fruit and stems is cut or shredded to approximately 0.1 mm to 10mm. The plant material is then ready for the extraction step.

[0032] To prepare the Tribulus terrestris L. extract, the preferablyshredded plant material is first mixed with water. A ratio ofapproximately 300 ml of deionized water for each 1 g of plant materialis preferable, but other ratios may be used. For example, 300 mldeionized (DI) water may be added to 1 g of plant material and blendedin a container for about 15 minutes to form a mixture. It is preferablethat sufficient speed be used to create a substantial vortex in themixture. Next, a solvent is added to the mixture, preferably in theratio of one part solvent to two parts of mixture. For example, about150 ml of a solvent such as ethyl alcohol may be added to 300 ml of themixture, mixed, and left for 48 hours, after which the mixture isfiltered. The solvent is then distilled off and the water extractconcentrated with a vacuum evaporator to about 1 ml for each g ofstarting mixture.

[0033] Next, the first extract is preferably treated to optimize thespirostanol content of the resulting composition. This is accomplishedby initially subjecting the first extract to a cleaving step, which ispreferably an enzymatic hydrolysis, but may also be acid hydrolysis, orother suitable methods. The hydrolysis step cleaves the furostanolscontained in the first extract, as illustrated in FIG. 8, to formbio-active spirostanols. The spirostanols are then separated andisolated from the first extract. This may be accomplished by usingchromatographic columns packed with silica gel or other packed materialto separate out spirostanols by size of the molecule using a mobilephase (organic solvent: alcohol:water) for each solvent quantity ratio1:100, wherein the first fractions to 10 are separated and added to thenext fresh solutions. The spirostanol saponins are contained in the next20 to 100 fractions.

[0034] All fractions are added together, dried in a vacuum evaporatorand purified two times with crystallization. The isolated spirostanolsare then added a second dried Tribulus extract that is prepared by, forexample, the ethyl alcohol extraction noted above, and blended.Alternately, spirostanols may be isolated by way of micro ultrafiltration, which separates out all active ingredients of the Tribulusterrestris extract by size of the molecule. The isolated spirostanolsare then preferably purified with crystallization and added to a seconddry extract to form an enriched extract. Using the method disclosedabove, the concentration of spirostanol saponins in the enriched dryextract may be from 0.1% to about 100%.

[0035] The enriched extract may be then be mixed with agents known inthe art to form, for example, a cream base, oral tablets, ointment orliquid, with an enriched extract concentration of preferably about 1% toabout 50% of the final composition, depending upon the therapeuticapplication. The enriched extract composition, however, may be increasedor decreased depending upon the therapeutic application. Any cream baseused for dermatological uses can be used. Alternatively, the extract maybe mixed with glycerin or paraffin to make a suppository form, or otheringredients known in the art to form oral tablets. Conventionalthickening agents, such as hydroxypropylmethylcellulose, may be used toadjust the viscosity to the degree best adapted to the form of thecomposition (solution, cream, gel etc.). Generally speaking, the presentinvention may be administered therapeutically by a vaginal cream, foam,ointment, or suppository. For advantageous application, the compositionin accordance with the invention can contain in addition, a bufferingagent such as hydrochloric acid in order to be able to adjust the pH toabout 4.7, which aids in maintaining healthy vaginal flora.

[0036] A preferred composition of the aqueous or enriched extract thatcan be used to treat vaginal infections, bacterial infections, viralinfections, such as herpes, acne and other conditions such as sunburn,hemorrhoids, diabetic skin conditions and phlebitis conditionspreferably comprises from about 70% to about 90% of the aqueous orenriched extract. A preferred cream or ointment may include from about70% to about 90% by weight or volume of Tribulus terrestris L aqueous orenriched extract; from about 1% to about 4% of a suitable humectant;from about 0.15% to about 2% of a preservative; from about 0.1% to about1% of a sequestering agent; from about 1% to about 5% of a thickener,suspending agent or stabilizer; from about 0.1% to about 4% of asoftening agent, and from about 0.1% to about 1% of a suitable solvent.Suitable humectants and preservatives include, respectively, glycerinand commercially available brands such as Germaben® II (propyleneglycol, diazolidinyl urea, methylparaben and propylparaben). Suitablesequestering agents include, for example, salts of ethylenediaminetetraacetic acid (e.g. sodium EDTA), and suitable thickeners includecommercial brands known in the art, such as Carbopol® products andCarbopol® 940. Suitable softening agents such as stearic acid andsuitable solvents such as triethanolamine may be used.

EXAMPLE I

[0037] Extraction of the plant material may be accomplished by themethod of the following example. Shred a mixture of leaves, fruit andstems of Tribulus Terrestris to a size of 0.1 to 10 mm in a shredder.For each 1 g of mixture, add about 300 ml deionized (DI) water andextract for about 15 min in a blender. Sufficient speed should be usedto create a substantial vortex. Add about 150 ml ethyl alcohol, mix,leave for 48 hours and then filter. Distill off the ethyl alcohol thenconcentrate the water extract with a vacuum evaporator to about 1 ml foreach g of starting mixture. Finally, mix with a cream base to aconcentration of 5% and 7.5% Tribulus Terrestris L concentrated waterextract. Any cream base used for dermatological uses can be used.Alternatively, mix with glycerin or paraffin to make into a suppositoryform.

EXAMPLE II

[0038] Creams made as described above with 5% and 7.5% TribulusTerrestris L enriched extract were used for local treatment in 275 womenwith vulvo-vaginitis. A physician carried out the treatment afterobtaining samples from the vagina, cervix and vulva. In most cases thecondition was classified as candidiasis. Application of the creams wasdone after the confirmation of the diagnosis through microbiologicaltesting. Quick improvement of the symptoms of the vulva was noted evenon the first day of the application of the cream.

[0039] The treatment had very good acceptance according to the opinionof the patients as well as their physicians. Vaginal antiseptic creamsare preparations for treatment of local problems of the vulva in theregion of the vulvo-vagina. The epithelia of the vagina is an effectivebarrier against infection, however, at the same time it is a propitiousenvironment for the development of protective flora and can permitdevelopment of numerous pathogenic microorganisms.

[0040] Their action has a maximal effectiveness in pH 4-5.6, whichprecisely the physiologic value of pH in the vagina as well as thepathological values of pH there.

[0041] Patients:

[0042] The investigation was conducted with women above 16 years of agewith vulvo-vaginitis with various etiology (bacteria, fungi, parasites,etc.) with at least two of the following symptoms: Leukorrhea, Pruritis,Burning, Edema and Erythema.

[0043] Patients with the associated symptoms of the upper genitalia wereexcluded from the study. Two hundred and seventy-five (275) patientswere followed for a period of 9 months. The patients' ages varied from16 to 62 years with a mean age of 32. More than 90% were younger than 45years. In 71.6% of the cases the patients were using contraception, mostoften oral (44.40%) and intrauterine (25.1%).

[0044] Lower genital pathology (vulvo-vaginitis) was the cause forconsultation in 63.3% of the cases and 29.1% were regular patients inwhom vulvo-vaginitis was diagnosed. A tendency of recurrence is seen inapproximately 40.7% of women diagnosed with vulvo-vaginitis. The initialdiagnosis during investigation was: candidiasis (83.9%); trichomonasis(7.3%); bacterial vaginosis (4.0%); chlamidia cervicitis (0.8%); herpes(0.8%); e-coli (0.8%); condyloma (0.8%); infected ectopia (0.8%);abscess of the left labia (0.8%); and diagnosis of mycosis was confirmedin 76% of the cases and rejected in 24%.

[0045] Treatment was carried out by applying Tribulus terristris L.cream as prepared according to the invention, to the whole vagina,including the cervix. Investigation consisted of three obligatorycheck-ups. The first check-up was done at the beginning of treatment(first day of the check-up) thorough clinical gynecologic check-up,obligatory microbiological investigation and competent endocervicalsampling. The second check up (second endocervical sample) was carriedout on the seventh day. The third check-up was done on the fourteenthday of the treatment. In cases of regression, without complete healingon the fourteenth day, a new application of Tribulus terrestris L. creamto the vagina was done.

[0046] The total effectiveness of the treatment was evaluated on thebasis of the development of the clinical symptoms, which motivated theperson to be included in the investigation: leukorrhea, pruritus,burning, edema and erythema. Each clinical symptom was evaluated on adegree from 0 to 3, according to the intensity during each check-up:0—lack; 1—of low importance; 2—important; and 3—very important.

[0047] The majority of important symptoms (No. 2 and 3) disappeared onthe second check-up on day 7 of the treatment (FIGS. 2 and 3). Resultswere evaluated on day 3, day 5 and day 7. In 93% of the cases thepatients were considered signifantly healed. That is the total result isless or equal on day 3, 5 and 7. Development of the symptoms wasevaluated each day by the patient up to the 7^(th) day of the treatment,which allowed for assessment of the speed of improvement of theleukorrhea, pruritus and burning. Symptoms of the leukorrhea, pruritusand burning had almost disappeared by the 3^(rd)-4^(th) day. See FIG. 4.

[0048] The cream was very well accepted by 9 out of 10 patients.Acceptance was perceived as good in 85% of the cases and mediocre in3.3% (FIG. 5). Subjectively, acceptance was considered good by 98.4% ofthe patients and bad by 2.4%.

EXAMPLE III

[0049] Clinical Investigation with Tribulus terrestris L. Acne Cream

[0050] 37 Patients with acne symptoms were treated with a cream-basedcomposition containing an enriched Tribulus terrestris L. extractprepared according to the method disclosed herein. The cream wasprepared by dissolving the dry extract of steroidal saponins enrichedwith spirostanol steroidal saponins, as disclosed herein, in purifiedwater. The mixture was then heated to 40-50° C. and added to the creambase, which is at a temperature of 60-65° C. The mixture is mixedcontinuously until the cream is homogenized. The concentration of dryenriched extract in the cream may be from approximately 0.1% to about50%, depending upon the therapeutic application. The concentration ofthe enriched spirostanol steroidal saponins in the dry extract, whichcontains steroidal saponins, sterols, and flavanoides isolated from theTribulus terrestris L. plant is preferably from about 0.1% to about 100%of the dry extract, depending upon the therapeutic application. Anointment can also be prepared, preferably at a concentration of fromabout 0.1% to about 50% of the enriched dry extract, and a suitablebase.

EXAMPLE IV

[0051] Preparation of Therapeutic Liquid for Eye and Ear Drops

[0052] Liquid Composition for Eye Infections

[0053] Dissolve 1% to 5% of the enriched extract prepared according tothe method disclosed herein in sterile purified water. The enrichedextract preferably has a spirostanol saponin content of about 0.1% toabout 100%, and may contain other sterols and bioflavanoides. Sodiumcitrate, citric acid, tyoxapol, edentate sodium may be added.

[0054] Liquid Composition for Ear Infections

[0055] Dissolve 1% to 50% of the enriched extract prepared according tothe method disclosed herein in sterile purified water. The enrichedextract preferably has a spirostanol saponin content of about 0.1% toabout 100%, and may contain other sterols and bioflavanoides. Sodiumcitrate, citric acid, tyoxapol , edentate sodium may be added.

[0056] The invention has been described with reference to particularembodiments. Other modifications and enhancements can be made withoutdeparting from the spirit and scope of the claims that follow.

EXAMPLE V

[0057] 305 Women, aged 40 to 67 years old (mean age, 50.9±5.1 years)were included in the study, which examined the effect of treatment withoral tablets, taken twice per day, containing 250 mg of Tribulusterrestris L. extract as prepared by the aqueous extraction process. Thesubjects were examined before taking the oral tablets and until at leastsix months after taking oral tablets. 83% of the subjects were

[0058] Each subject was interviewed as to qualitative factors such asthe frequency and severity of hot flushes, parenthesis, sleepdisturbances, irritability, depression etc. A qualitative average of thesymptoms was prepared, using a scale from K=0 to K=3, with K=0 being nosymptoms, K=1 being light symptoms not influencing the quality of life,K=2 being moderate symptoms affecting the quality of life and K=3 beingintense symptoms seriously affecting the quality of life. The mean foreach symptom for the study group (n=305) is illustrated in Table 1,below: TABLE 1 Mean Severity of Symptoms STATISTICAL DATA PERCENT OFSTUDY SYMPTOM (K) (MEAN SEVERITY) SUBJECTS (N = 305) Hot flushes 3 96%(291) Parenthesis 2 44% (133) Sleep disturbances 2 77% (235)Irritability 2 76% (230) Depression 1 59% (179) Asthenia 1 66% (200)Artalga/mialgia 1 52% (158) Headache 1 46% (141) Shivers 1 37% (112)Dizziness 1 28% (86)  Palpitations 1 36% (110)

[0059] A quantitative assessment for menopausal symptoms is generallycarried out with the help of Kuppermann Menopause Index. Typicalsymptoms, such as those listed in Table 1, are classified and gradedaccording to a corresponding multiplication constant. This constant isrespectively multiplied by the subjective grading factor (from 0 to 3),as discussed in reference to Table 1. The resulting value are then addedand the total score used as an indication for the severity of theclimacteric syndrome (menopause). A score of 21-35 is moderate toaverage and a score of 15-20 points indicates a low degree of severity.A score higher than 35 points indicates severe symptoms. During thecourse of treatment, the efficacy of a treatment may be established byrecording the decrease in the Kuppermann index.

[0060] 48% of the subjects (147 of n=305) were treated with the herbalextract as described above. 95.4% of subjects (291 of n=304) complainedof hot flushes; with 27% of subjects describing mild symptoms (K=1) in27%, and 69% describing moderate or intense (K=2-3) symptoms. 66% ofsubjects (200 of n=305) indicated that vulvo-vaginal trophy created mildsymptoms (K=1). Other frequent symptoms were sleep disturbances,depression and fatigue. The Kaupermann Index ranged from 0-50 for thestudy group, with a median index of 20.0±9.6. 6 subjects had anomaliesfound during mammal gland examination and 10 subjects had anomalies ofthe thyroid gland. 1 subject had a prolapsed uterus. One out of threesubjects displayed symptoms of venous insufficiency or varicose veins ofthe lower extremities. 24 subjects were removed from the study for thefollowing reasons: loss of vision (3); voluntary removal (5); pathologydiscovered during study (2); intolerance to therapy (2); ineffectivenessof therapy (8); non-medical reasons (4).

[0061] After six months of treatment, the percentage of subjectscomplaining of hot flushes fell to 11.5% (26 out of n=226). The meanindex of Kuppermann fell from 20±9.6 at M=0 to 4.3±5.6 at M=6.Improvements in the skin, epidermal and vulvo-vaginal trophy were alsonoted. The treatment was effective in improving symptoms in 90% of thesubjects.

[0062] During the study period, a minimal increase in mean body weightof 600 g was noted by M=3 (mean weight=60.2±8.3 Kg (n=218)), whichleveled after M=3 (mean weight 60.1±8.3 Kg) through the end of thestudy. The pre-study weight of the subjects ranged from 40 to 90 Kg,with a mean of 59.8±8.5 Kg. No significant changes in systolic bloodpressure were noted during the study period(mean of 125.5 mm Hg at M=0,n=222). Diastolic blood pressure increased slightly (up 1.8 mm Hg) fromM=0 to M=3 and leveled after M=3 to the end of the study. The M=0diastolic pressure was measured at 75 mm Hg±8.6 mm Hg; 76.8±8.3 at M=3;and 75.9±8.5 mm Hg at M=6. A correlation of the beneficence/risk of thetherapy showed that therapy was well tolerated by 77% of subjects (218of n=284) (FIG. 11). 70% of the subjects (202 of n=287) had a positiveor very positive assessement of their condition after treatment.

EXAMPLE VI

[0063] The patient is a 48-year old male, previous intravenous drugabuser, diagnosed with Hepatitis-C, in November 1999. He was asymptomatic. The diagnosis was prompted by an abnormal liver functiontest, on a routine yearly health evaluation. At the time of evaluation,he had been off illicit drugs for at least five years.

[0064] Laboratory tests revealed elevated liver enzymes. Subsequentscreening for hepatitis B and C was positive for the B surface antibodyand HCV antibody. The hepatitis C RNA quantitative test by the Chironmethod done upon diagnosis in 1999 was 2,650,000 IU/ml. A repeatquantitative hepatitis C viral RNA by PCR done approximately one yearlater was 392,000 IU/ml.

[0065] Approximately another year later, about twenty-two monthspost-diagnosis, HCV RNA quantitative by PCR method revealed a vital loadof >850,000; with a sensitivity limit of 300,000-850,000. At this point,the patient began orally taking tablets comprising extract of Tribulusterrestris L. About another ten (10) months later, after treatment withTribulus began, the HCU RNA quantitative by PCR method revealed a viralload of 334,000 with a sensitivity limit of 300,000 to 850,000 (IU/ml).

What is claimed is:
 1. An antibacterial, anti-viral, anti-fungaltherapeutic composition for topical treatment of an infection,comprising an extract of Tribulus terristris L.
 2. The composition ofclaim 1, wherein the Tribulus terristris L comprises a first Tribulusterrestris L. extract and wherein the first Tribulus terristris L.extract is treated to convert furostanol saponin compounds containedtherein to spirostanol compounds.
 3. The composition of claim 2, whereinthe first extract is subjected to a step that cleaves furostanolcompounds contained in the first extract to form spirostanol compounds.4. The composition of claim 3, wherein the spirostanol compounds areisolated from a remaining portion of the first extract and are added toa second Tribulus terristris L. extract to form an enriched extract. 5.The composition of claim 4, wherein the spirostanol compounds areisolated from a remaining portion of the first extract by a methodselected from the group consisting of chromatographic methods andmicro-filtration methods.
 6. The composition of claim 2, wherein thepercentage by weight of spirostanol compounds in the therapeuticcomposition is from approximately 0.1% to approximately 100%.
 7. Thecomposition of claim 6, wherein the percentage of spirostanol compoundsin the therapeutic composition is from approximately 20% toapproximately 100%.
 8. The composition of claim 7, wherein thepercentage of spirostanol compounds in the therapeutic composition isfrom approximately 50% to approximately 80%.
 9. The composition of claim2 comprising: a. from about 70% to about 90% by weight of TribulusTerrestris L extract; b. from about 1% to about 4% of a humectant; c.from about 0.15% to about 2% of a preservative; d. from about 0.1% toabout 1% of a sequestering agent; e. from about 1% to about 5% of anagent selected from the group consisting of thickening agents,suspension agents and stabilizing agents; f. from about 1% to about 4%of a softening agent; and g. from about 0.1% to about 1% of a solvent.10. The composition of claim 9 wherein the humectant comprises glycerin.11. The composition of claim 9, wherein the softening agent comprisesstearic acid.
 12. The composition of claim 9, wherein the solventcomprises triethanolamine.
 13. The composition of claim 9, wherein thesequestering agent comprises ethylenediamine tetraacetic acid.
 14. Amethod to treat an infection caused by an organism selected from thegroup consisting of bacteria, viruses, parasites and fungi, comprisingthe steps of: a. applying a topical medication comprising a firstextract of Tribulus terrestris L to an affected area of a patient. 15.The method of claim 14, comprising the further step of treating thefirst extract to convert furostanol saponin compounds contained thereinto spirostanol compounds before applying the therapeutic composition tothe affected area of a patent.
 16. The method of claim 15 comprising thefurther steps of: a. isolating the spirostanol compounds from the firstextract after treating the first extract to convert furostanol compoundsa contained therein to spirostanol compounds; and b. adding the isolatedspirostanol compounds to a second Tribulus terrestris L. extract to forman enriched extract.
 17. The method of claim 16 comprising the furthersteps of: a. adding the enriched extract to a base suitable for mucosalapplication to form the topical medication; and b. applying the topicalmedication to the affected area of a patient.
 18. The method of claim17, wherein the mucosal tissue comprises vaginal tissue.
 19. The methodof claim 18, wherein the organism is selected from the group consistingof candidiasis, trichomanasis, chlamidia, herpes, e-coli, condyloma andmyycogens
 20. The method of claim 19, wherein the topical medicationcomprises from about 5% to about 7.5% of the enriched extract.
 21. Themethod of claim 17, wherein the mucosal tissue comprises eye tissue. 22.The method of claim 21, wherein the topical medication comprises fromabout 1% to about 5% of the enriched extract.
 23. A method to treat anarea of the skin affected by acne, comprising the steps: a. preparing anaqueous first extract of Tribulus terrestris L; b. treating the firstextract to convert the furostanol saponin compounds contained therein tospirostanol compounds; c. isolating the spirostanol compounds from thefirst extract; d. adding the isolated spirostanol compounds to a secondTribulus terrestris L. extract to form an enriched extract; e. addingthe enriched extract to a base suitable for application to the skin toform a topical medication; and f. applying the topical medication to theaffected area.
 24. The method of claim 23, wherein the topicalmedication comprises between about 0.1% to about 100% of the enrichedextract.
 25. A method of testing and preparing for readiness ofharvesting tribulus Terrestris L. comprising the steps of: a. collectinga sample of leaves, fruit and stems of Tribulus Terrestris L; b. addingabout one part water to about 2.5 parts of the sample; c. blending forabout 15 minutes in a blender with sufficient speed to create a vortex;d. immediately examining the level of foam and water in the blender; ande. if there is at least 1.5 times as much foam as water, harvesting theTribulus Terrestris L. within the week, otherwise repeating steps a.through e. at a later time.